This invention relates to novel xcex1-sulfonamido and xcex1-sulfinamido containing carboxylic acid compounds which are potent inhibitors of Arginyl-Glycyl-Aspartyl-(RGD)-dependent integrins. In another aspect, the present invention relates to these xcex1-sulfonamido and xcex1-sulfinamido containing carboxylic acids, their pharmaceutically-acceptable salts, and pharmaceutically-acceptable compositions thereof which are useful as potent inhibitors of integrin adhesive functions in mammals. In yet another aspect, the invention relates to methods for using these inhibitors as therapeutic agents for disease states in mammals characterized by aberrant cellular adhesive disorders that occur during thrombosis and restenosis.
Cellular adhesion is believed to play an important role in both thrombus formation and cellular responses to vascular injury, as well as for normal hemostasis. Vascular injury and thrombosis are prevalent during the development and progression of vascular disease states. These include conditions such as atherosclerosis, acute myocardial infarction, chronic stable angina, unstable angina, transient ischemic attacks, stroke, peripheral vascular disease, arterial thrombosis, and conditions induced by interventional procedures such as restenosis following angioplasty.
Cellular adhesions can be characterized as either cell-cell adhesions or cell-matrix adhesions. Cells utilize a variety of cell surface adhesion receptors and adhesive proteins to facilitate these adhesive interactions. For cell-cell type adhesions, platelets play a major role in this type of adhesive interaction that occurs during acute thrombosis. Platelet aggregation, thrombus formation and consolidation of clots mediated by platelets are principally achieved by adhesive protein crosslinking of the platelet glycoprotein (GPIIb-IIIa) also referred to as xcex1IIxcex2xcex23 which is found on the platelet surface. This heterodimeric adhesion receptor is one member of a large family of heterodimeric transmembrane glycoprotein receptors, called integrins (Hynes, R. O., xe2x80x9cIntegrins: Versatility, Modulation and Signaling in Cell Adhesionxe2x80x9d, Cell 69:11 (1992)).
Other integrins which may have important cell adhesion functions in thrombosis, hemostasis or in disease states characterized by vascular injury are the vitronectin receptors (xcex1vxcex23 and xcex1vxcex25) and the fibronectin receptor (xcex15xcex21) In particular, the vitronectin receptor, xcex1vxcex23, has been postulated to play roles in cellular migration of smooth muscle cells following vascular injury that can ultimately lead to restenosis of the vessel (Yue, T. L., et at., xe2x80x9cOsteopontin-Stimulated Vascular Smooth Muscle Cell Migration is Mediated by xcex23 Integrinxe2x80x9d, Exp. Cell. Res. 214:459-464 (1994); Choi, E. T., et al., xe2x80x9cInhibition of Neointimal Hyperplasia by Blocking xcex1vxcex23 Integrin with a Small Peptide Antagonist GpenGRGDSPCAxe2x80x9d, J. Vasc. Surg. 19:125-134 (1994); Matsuno, H., xe2x80x9cInhibition of Integrin Function by A Cyclic RGD-Containing Peptide Prevents Neointima Formationxe2x80x9d, Circulation 90:2203-2206 (1994)). A number of the natural ligands of these integrins (e.g. xcex1IIxcex2xcex23, xcex1vxcex23, xcex1vxcex25, and xcex15xcex21) such as fibrinogen, fibronectin, von Willebrand factor, thrombospondin, osteopontin, vitronectin and others, contain and utilize the tripeptide sequence, Arg-Gly-Asp (RGD) to bind to their respective integrins. Small synthetic peptides containing the RGD sequence have been shown to bind to these integrins and to compete for the binding of natural adhesive ligands (Rouslahti, E. and Pierschbacher, M. D.,xe2x80x9cNew Perspectives in Cell Adhesion: RGD and Integrinsxe2x80x9d, Science 238:491-497 (1987)). Peptides containing the RGD sequence or mimetic compounds have thus been the basis for the discovery of several potent and highly specific inhibitors of platelet xcex1IIxcex2xcex23 which are useful as antithrombotic agents. This literature has been extensively reviewed. See Coller, B. S., xe2x80x9cBlockade of Platelet GPIIb/IIIa Receptors as an Antithrombotic Strategyxe2x80x9d, Circulation 92:2373-2380 (1995); Cook, N. S., et al., xe2x80x9cPlatelet Glycoprotein IIb/IIIa Antagonistsxe2x80x9d, Drugs of the Future 19:135-159 (1994); T. Weller., et al., xe2x80x9cFibrinogen Receptor Antagonistsxe2x80x94A Novel Class of Promising Antithromboticsxe2x80x9d, Drugs of the Future 19:461 (1994); and Zablocki, J. A., et al., xe2x80x9cFibrinogen Receptor Antagonistsxe2x80x9d, Exp. Opin. Invest. Drugs, 3:437-448 (1994).
Highly specific inhibitors of xcex1vxcex23 based on the RGD recognition sequence have also been recently described. Specifically, the cyclic peptide, cyclo[Arg-Gly-Asp-D-Phe-Val] is a very potent and specific inhibitor of the vitronectin receptor xcex1vxcex23 (Pfaff, M., et al., xe2x80x9cSelective Recognition of Cyclic RGD Peptides of NMR Defined Conformation by xcex1IIxcex2xcex23, and xcex15xcex21 Integrinsxe2x80x9d, J. Biol. Chem. 269:20233-20238 (1994); Jonczyk, A., et al., European Patent Application 578083A2 (1994)).
The present invention describes the preparation of novel compounds which inhibit the adhesive function of various RGD-dependent integrins. More specifically, the novel compounds are non-specific inhibitors of the platelet integrin xcex1IIxcex2xcex23 and the vitronectin receptor xcex1vxcex23.
The present invention relates to novel xcex1-sulfonamido and xcex1-sulfinamido containing carboxylic acids or carboxylic esters, their pharmaceutically-acceptable .stereoisomers, salts, hydrates, solvates and prodrug derivatives, and pharmaceutically-acceptable compositions thereof which have particular biological properties and are useful as potent antithrombotics and/or antirestenotic agents in mammals.
The present invention provides a compound of the formula: 
wherein:
Y is selected from the group consisting of xe2x80x94COOH, xe2x80x94PO3H2, xe2x80x94SO3H and xe2x80x94COOR4; where R4 is selected from the group consisting of C1-10alkyl, C1-8alkylaryl, aryl-C1-8alkyl, C1-8alkyloxycarbonyloxy-C1-8alkyl, aryloxycarbonyloxy-C1-8alkyl,
C1-8alkyloxycarbonyloxyaryl, C1-8alkylcarbonyloxy-C1-8alkyl, arylcarbonyloxy-C1-8alkyl and C1-8alkylcarbonyloxyaryl;
A is selected from the group consisting of C6-12alkyl, C0-8alkyl-NR5xe2x80x94COxe2x80x94C0-8alkyl, C0-8-alkyl-COxe2x80x94NR5xe2x80x94CO0-t8alkyl, C0-8alkyl-Oxe2x80x94C0-8-alkyl, C0-8alkyl-NR5xe2x80x94COxe2x80x94C0-8alkyl-NR5-COxe2x80x94C0-8alkyl, C0-8alkyl-NR5xe2x80x94COxe2x80x94C1-8alkyl-COxe2x80x94NR5xe2x80x94C0-8alkyl, C0-8alkyl-COxe2x80x94NR5-C1-8alkyl-NR5xe2x80x94COxe2x80x94C0-8alkyl, C0-8alkyl-COxe2x80x94NR5xe2x80x94C0-8alkyl-COxe2x80x94NR5xe2x80x94CO-alkyl, Cll8alkyl-COxe2x80x94C1-8alkyl-COxe2x80x94NR5xe2x80x94C0-8alkyl, C0-8alkyl-COxe2x80x94C0-8alkyl-NR5xe2x80x94COxe2x80x94C0-8alkyl, C0-8alkyl-Oxe2x80x94C2-8alkyl-NR5xe2x80x94COxe2x80x94C0-8alkyl, C0-8alkyl-Oxe2x80x94C0-8alkyl-COxe2x80x94NR5xe2x80x94C0-8alkyl, C0-8alkyl-Oxe2x80x94C2-8alkyl-Oxe2x80x94C0-8alkyl-COxe2x80x94NR5-CO-alkyl, C0-8alkyl-Sxe2x80x94C0-8alkyl, C0-8alkyl-S(On)-C0-8alkyl, C0-8alkyl-Sxe2x80x94C2-8alkyl-NR5xe2x80x94COxe2x80x94C0-8alkyl, C0-8alkyl-S(On)-C2-8alkyl-NR5xe2x80x94COxe2x80x94C0-8alkyl, C0-8alkyl-Sxe2x80x94C0-8alkyl-COxe2x80x94NR5xe2x80x94C0-8alkyl, C0-8alkyl-S(On)-C1-8-alkyl-COxe2x80x94NR5xe2x80x94C0-8alkyl, C0-8alkyl-NR5xe2x80x94COxe2x80x94C0-8alkyl-Sxe2x80x94C0-8alkyl, C0-8alkyl-NR5xe2x80x94COxe2x80x94C0-8alkyl-S(On)-C1-8alkyl, C0-8alkyl-COxe2x80x94NR5-C2-8alkyl-Sxe2x80x94C0-8-alkyl, C0-8alkyl-COxe2x80x94NR5-C2-8alkyl-S(On)-C0-8alkyl, C0-8alkyl-NR5-C0-8alkyl-CO2xe2x80x94C0-8alkyl, C0-8alkyl-NR5-C0-8alkyl-CS-Oxe2x80x94C0-8alkyl, C0-8alkyl-NR5-C0-8alkyl-COxe2x80x94NR5-C0-8alkyl, C0-8alkyl-NR5-C0-8alkyl-CS-NR5xe2x80x94C0-8alkyl, C0-8alkyl-Oxe2x80x94C0-8alkyl-CO2xe2x80x94C0-8alkyl, C0-8alkyl-Oxe2x80x94C0-8alkyl-CS-Oxe2x80x94C0-8alkyl; C0-8alkyl-SiR7R8-alkyl, C0-8alkyl-SiR7R8xe2x80x94C0-8alkyl-NR6xe2x80x94COxe2x80x94C0-8alkyl, and C0-8alkyl-SiR7R8-C0-8alkyl-COxe2x80x94NRxe2x80x94CO-alkyl; where R5, R6, R7 and R8 are independently selected from the group consisting of H and C 1-6alkyl; and where n=1 or 2;
Z is selected from the group consisting of xe2x80x94NHxe2x80x94C(NR9R10)xe2x95x90NR11, xe2x80x94NHxe2x80x94C(R9)xe2x95x90NR11, xe2x80x94C(NR9R10)xe2x95x90NR11 and piperidinyl; where R9, R10 and R11 are independently selected from the group consisting of H, C1-6alkyl, aryl-C1-3alkyl and aryl; or where two of the R9, R10 or R11 substituents form a cyclic ring containing (CH2)p, where p=2-5;
R1 is H;
R2 is selected from the group consisting of xe2x80x94SOm-aryl, xe2x80x94SOmxe2x80x94C1-10alkyl and xe2x80x94SOm-heteroaryl, where m=1-2;
R3 is selected from the group consisting of H, C1-8alkyl, aryl, C1-8alkylaryl and heteroaryl;
and all pharmaceutically-acceptable stereoisomers, salts, hydrates, solvates and prodrug derivatives thereof.
In certain aspects of this invention, compounds are provided which are useful as diagnostic reagents. In another aspect, the present invention includes pharmaceutical compositions comprising a pharmaceutically-effective amount of the compounds of this invention and a pharmaceutically-acceptable carrier. In yet another aspect, the present invention includes methods which comprise administering the compounds of the present invention and pharmaceutical compositions thereof for preventing or treating disease states characterized by thrombosis or vascular injury in mammals. Optionally, the methods of this invention comprise administering such pharmaceutical compositions in combination with an additional therapeutic agent such as an antithrombotic, a thrombolytic agent or an anticoagulant, or any combination thereof.
The preferred compounds also include their pharmaceutically-acceptable stereoisomers, hydrates, solvates, salts and prodrug derivatives.
In accordance with the present invention and as used herein, the following terms are defined with the following meanings, unless explicitly stated otherwise.
The term xe2x80x9calkylxe2x80x9d refers to saturated and unsaturated aliphatic groups including straight-chain and branched-chain and cyclic groups, or any combination thereof, having the number of carbon atoms specified, or if no number is specified, having up to 12 carbon atoms. Cyclic alkyls typically comprise a monocyclic aliphatic ring having 3 to 12 carbon atoms and preferably 3 to 7 carbon atoms. The cyclic alkyls of this invention may include one or more nitrogen atoms. Preferably, xe2x80x9calkylxe2x80x9d refers to straight-chain and branched-chain groups; more preferably straight-chain groups.
The term xe2x80x9carylxe2x80x9d refers to an unsubstituted or substituted aromatic ring, substituted with one, two or three substituents selected from C1-6alkoxy, C1-6alkyl, C1-6alkylamino, hydroxy, halogen, cyano, hydroxyl, mercapto, nitro, thioalkoxy, carboxaldehyde, carboxyl, carboalkoxy and carboxamide, including but not limited to carbocyclic aryl, heterocyclic aryl, and biaryl groups and the like, all of which may be optionally substituted. Preferred aryl groups include phenyl, halophenyl, C1-6alkylphenyl, naphthyl, biphenyl, phenanthrenyl, naphthacenyl, and aromatic heterocyclics. The term xe2x80x9cheteroarylxe2x80x9d as used herein refers to any aryl group, containing from one to four heteroatoms, selected from the group consisting of nitrogen, oxygen and sulfur.
The term xe2x80x9carylalkylxe2x80x9d refers to one, two, or three aryl groups appended to an alkyl group having the number of carbon atoms designated. Suitable arylalkyl groups include, but are not limited to, benzyl, picolyl, naphthylmethyl, phenethyl, benzhydryl, trityl, and the like, all of which may be optionally substituted. Similarly, the term xe2x80x9calkylarylxe2x80x9d refers to an alkyl group, having the number of carbon atoms designated, appended to one, two, or three aryl groups.
The terms xe2x80x9chaloxe2x80x9d or xe2x80x9chalogenxe2x80x9d as used herein refer to Cl, Br, F or I substituents.
The term xe2x80x9coxyxe2x80x9d refers to an oxygen (O) atom. The terms xe2x80x9calkyloxyxe2x80x9d and xe2x80x9caryloxyxe2x80x9d thus refer to the respective groups positioned adjacent to an oxygen atom. The term carbonyloxy refers to xe2x80x94C(O)xe2x80x94Oxe2x80x94.
The term xe2x80x9cpharmaceutically-acceptable saltsxe2x80x9d includes salts of compounds derived from the combination of a compound of the present invention and an organic or inorganic acid. These compounds are useful in both free base and salt form. In practice, the use of the salt form amounts to use of the base form; both acid and base addition salts are within the scope of the present invention.
xe2x80x9cPharmaceutically-acceptable acid addition saltxe2x80x9d refers to those salts which retain the biological effectiveness and properties of the free bases and which are not biologically or otherwise undesirable, formed with inorganic acids such as hydrochloric acid, hydrobromic acid, hyroiodic acid, sulfuric acid, nitric acid, phosphoric acid and the like, and organic acids such as acetic acid, propionic acid, glycolic acid, pyruvic acid, oxalic acid, maleic acid, malonic acid, succinic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, salicylic acid and the like.
xe2x80x9cPharmaceutically-acceptable base addition saltsxe2x80x9d include those derived from inorganic bases such as sodium, potassium, lithium, ammonium, calcium, magnesium, iron, zinc, copper, manganese, aluminum salts and the like. Particularly preferred are the ammonium, potassium, sodium, calcium and magnesium salts. Salts derived from pharmaceutically-acceptable organic nontoxic bases include salts of primary, secondary, and tertiary amines, substituted amines including naturally occurring substituted amines, cyclic amines and basic ion-exchange resins, such as isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, ethano,amine, 2-diethylaminoethanol, trimethamine, dicyclohexylamine, lysine, arginine, histidine, caffeine, procaine, hydrabamine, choline, betaine, ethylenediamine, glucosamine, methylglucamine, theobromine, purines, piperizine, piperidine, N-ethylpiperidine, polyamine resins and the like. Particularly preferred organic nontoxic bases are isopropylamine, diethylamine, ethanolamine, trimethamine, dicyclohexylamine, choline, and caffeine.
The term xe2x80x9cprodrug derivativesxe2x80x9d refers to compounds of the invention which have metabolically cleavable groups and become, by sovolysis or under physiological conditions, compounds of the invention which are pharmaceutically-active in vivo. Fro example, ester derivatives of compounds of this invention are often active in vivo, but may have only weak or no activity in vitro. Other derivatives of the compounds of this invention have activity in both their acid and acid derivative forms, but the acid derivative form often offers advantages of solubility, tissue compatibility, or delayed release in the mammalian organism. See Bundgard, H., xe2x80x9cDesign of Prodrugsxe2x80x9d, pp. 7-9, 21-24, Elsevier, Amsterdam, 1985. Prodrugs include acid derivatives well known to practitioners of the art, such as esters prepared by the reaction of the parent acid with a suitable alcohol, or amides prepared by reaction of the parent acid with an amine. Simple aliphatic or aromatic esters derived from acidic groups pendant on the compounds of this invention are preferred prodrug derivatives. In some cases, it is desirable to prepare double ester-type prodrugs such as (aclyoxy)alkyl esters or [(alkoxycarbonyl)oxy]alkyl esters.
xe2x80x9cBiological propertyxe2x80x9d for the purposes herein means an in vivo effector or antigenic function or activity that is directly or indirectly performed by a compound of this invention. Effector functions include receptor or ligand binding, any enzyme activity or enzyme modulatory activity, any carrier binding activity, any hormonal activity, any activity in promoting or inhibiting adhesion of cells to an extracellular matrix or cell surface molecules, or any structural role. Antigenic functions include possession of an epitope or antigenic site that is capable of reacting with antibodies raised against it.
In addition, the following are used in this application:
xe2x80x9cBocxe2x80x9d t-butoxycarbonyl
xe2x80x9cBOPxe2x80x9d benzotriazol-1-yloxy-tris-(dimethylamino)phosphonium hexafluorophosphate
xe2x80x9cCbzxe2x80x9d benzyloxycarbonyl
xe2x80x9cDCCxe2x80x9d N,Nxe2x80x2-dicyclohexylcarbodiimide
xe2x80x9cDIEAxe2x80x9d diisopropylethylamine
xe2x80x9cDMAPxe2x80x9d 4-dimethylaminopyridine
xe2x80x9cDMFxe2x80x9d N,N-dimethylformamide
xe2x80x9cHBTUxe2x80x9d 2-(1-H-benzotriazol-1-yl)-1,1,3,3,-tetramethyluronium hexafluorophosphate
xe2x80x9cHFxe2x80x9d hydrogen fluoride
xe2x80x9cHOBtxe2x80x9d N-hydroxybenzotriazole
xe2x80x9cMeOHxe2x80x9d methanol
xe2x80x9cPhxe2x80x9d phenyl
xe2x80x9cPhSO2NHxe2x80x94xe2x80x9d phenylsulfonamido
xe2x80x9cTFAxe2x80x9d trifluoroacetic acid
In the compounds of this invention, carbon atoms bonded to four non-identical substituents are asymmetric. Accordingly, the compounds may exist as stereoisomers, including enantiomers and diastereomers. The compounds of this invention having one or more centers of asymmetry may exist as enantiomers or mixtures thereof (e.g. racemates). In addition, compounds that have two or more asymmetric centers can exist as diastereomers. The syntheses described herein may employ racemates, enantiomers or diastereomers as starting materials or intermediates. Diastereomeric products resulting from such syntheses may be separated by chromatographic or crystallization methods, or by other methods known in the art. Likewise, enantiomeric product mixtures may be separated using methods known in the art. See for example, Jacqes, Collet and Wilen xe2x80x9cEnantiomers, Racemates and Resolutionsxe2x80x9d (Krieger Publishing Co., Malabar, Fla. 1991). Each of the asymmetric centers, when present in the compounds of this invention, may be in one of two configurations (R or S), and both are within the scope of the present invention. Some of the compounds may be designated either D or L, which is a less preferred indicator of the configuration of the compound, based on the compound of this invention having a configuration that is similar to known amino acids. In the processes described above, the final products may, in some cases, contain a small amount of the products having D or L-form residues; however, these products do not affect their therapeutic or diagnostic application.
In all of the compounds of the invention having one or more amide linkages (xe2x80x94COxe2x80x94NHxe2x80x94), such amide linkages may optionally be replaced with another linkage which is an isostere such as xe2x80x94CH2NHxe2x80x94, xe2x80x94CH2Sxe2x80x94, CH2xe2x80x94O, CH2CH2, xe2x80x94CHxe2x95x90CHxe2x80x94 (cis and trans), xe2x80x94COCH2xe2x80x94, xe2x80x94CH(OH)CH2xe2x80x94, xe2x80x94CH2SOxe2x80x94, and CH2SO2. This replacement can be made by methods known in the art. The following references describe preparation of peptide analogs which include these alternative-linking moieties: Spatola, A. F., Vega Data (March 1983), Vol. 1, Issue 3, xe2x80x9cPeptide Backbone Modificationsxe2x80x9d (general review); Spatola, A. F., in xe2x80x9cChemistry and Biochemistry of Amino Acids, Peptides and Proteins,xe2x80x9d B. Weinstein, eds., Marcel Dekker, New York, p. 267 (1983) (general review); Morley, J. S., Trends Pharm Sci pp. 463-468 (1980) (general review); Hudson, D., et al., Int J Pept Prot Res 14:177-185 (1979) (xe2x80x94CH2NHxe2x80x94, xe2x80x94CH2CH2xe2x80x94); Spatola, A. F., et al., Life Sci 38:1243-1249(1986) (xe2x80x94CH2xe2x80x94S); Hann, M. M., J Chem Soc Perkin Trans I pp. 307-314 (1982) (xe2x80x94CHxe2x95x90CHxe2x80x94, cis and trans); Almquist, R. G., et al., J Med Chem 23:1392-1398 (1980) (xe2x80x94COCH2xe2x80x94); Jennings-White, C., et al., Tetrahedron Lett 23:2533(1982) (xe2x80x94COCH2xe2x80x94); Szelke, M., et al., European Application EP 45665; CA 97:39405 (1982) (xe2x80x94CH(OH)CH2xe2x80x94); Holladay, M. W., et al., Tetrahedron Lett 24:4401-4404 (1983) (xe2x80x94C(OH)CH2xe2x80x94); and Hruby, V. J., Life Sci 31:189-199 (1982) (xe2x80x94CH2xe2x80x94Sxe2x80x94).
In preferred embodiments, the present invention provides compounds of the formula: 
wherein:
Y is selected from the group consisting of xe2x80x94COOH, xe2x80x94PO3H2, xe2x80x94SO3H and xe2x80x94COOR4; where R4 is selected from the group consisting of C1-10alkyl, C1-8alkylaryl, aryl-C1-8alkyl, C1-8alkyloxycarbonyloxy-C1-8alkyl, aryloxycarbonyloxy-C1-8alkyl, C1-8alkyloxycarbonyl-oxyaryl, C1-8alkylcarbonyloxy-C1-8alkyl, arylcarbonyloxy-C1-8alkyl and C1-8alkylcarbonyl-oxyaryl;
A is selected from the group consisting of C6-12alkyl, C0-8alkyl-NR5xe2x80x94COxe2x80x94C0-8alkyl, C0-8alkyl-COxe2x80x94NR5-C0-8alkyl, C0-8alkyl-Oxe2x80x94C0-8alkyl, C0-8alkyl-NR5xe2x80x94COxe2x80x94C1-8alkyl-NR5xe2x80x94COxe2x80x94C0-8alkyl, C0-8alkyl-NR5xe2x80x94COxe2x80x94C1-8alkyl-COxe2x80x94NR5xe2x80x94C0-8-alkyl, C0-8alkyl-COxe2x80x94NR5-C1-8alkyl-NR5xe2x80x94COxe2x80x94C0-8alkyl, C0-8alkyl-COxe2x80x94NR5xe2x80x94C0-8alkyl-COxe2x80x94NR5xe2x80x94C0-8alkyl, C0-8alkyl-COxe2x80x94C1-8alkyl-COxe2x80x94NR5xe2x80x94C0-8alkyl, C0-8alkyl-COxe2x80x94C0-8alkyl-NR5xe2x80x94COxe2x80x94C0-8-alkyl, C0-8alkyl-Oxe2x80x94C2-8alkyl-NR5xe2x80x94COxe2x80x94C0-8alkyl, C0-8alkyl-Oxe2x80x94C0-8alkyl-COxe2x80x94NR5-C0-8alkyl, C0-8alkyl-Oxe2x80x94C2-8alkyl-Oxe2x80x94C0-8alkyl-COxe2x80x94NR5xe2x80x94C0-8alkyl, C0-8alkyl-Sxe2x80x94C0-8alkyl, C0-8alkyl-S(On)xe2x80x94C0-8alkyl, C0-8alkyl-Sxe2x80x94C2-8alkyl-NR5xe2x80x94COxe2x80x94C0-8alkyl, C0-8alkyl-S(On)xe2x80x94C2-8alkyl-NR5xe2x80x94COxe2x80x94C0-8alkyl, C0-8alkyl-Sxe2x80x94C0-8alkyl-COxe2x80x94NR5xe2x80x94C0-8alkyl, C0-8alkyl-S(On)xe2x80x94C1-8alkyl-COxe2x80x94NR5xe2x80x94C0-8alkyl, C0-8alkyl-NR5xe2x80x94COxe2x80x94C0-8alkyl-Sxe2x80x94C0-8alkyl, C0-8alkyl-NR5xe2x80x94COxe2x80x94C1-8alkyl-S(On)xe2x80x94C0-8alkyl, C0-8alkyl-COxe2x80x94NR5-C2-8alkyl-Sxe2x80x94C0-8alkyl, C0-8alkyl-COxe2x80x94NR-C2-8alkyl-S(On)-C0-8alkyl, C0-8alkyl-NR5xe2x80x94C0-8alkyl-CO2-C0-8alkyl, C0-8alkyl-NR5-C0-8alkyl-CSxe2x80x94Oxe2x80x94C0-8alkyl, C0-8alkyl-NR5-C0-8alkyl-COxe2x80x94NR5-C0-8alkyl, C0-8alkyl-NR5-C0-8alkyl-CSxe2x80x94NR5-C0-8alkyl, C0-8alkyl-Oxe2x80x94C0-8alkyl-CO2-C0-8alkyl, C0-8alkyl-Oxe2x80x94C0-8-alkyl-CSxe2x80x94Oxe2x80x94C0-8-alkyl; C0-8alkyl-SiR7R8-C0-8alkyl, C0-8alkyl-SiR7R8-C0-8alkyl-NR6xe2x80x94COxe2x80x94C0-8alkyl, and C0-8alkyl-SiR7R8-C0-8alkyl-COxe2x80x94NR6-C0-8alkyl; where R5, R6, R7 and R8 are independently selected from the group consisting of H and C1-6alkyl; and where n=1 or 2;
Z is selected from the group consisting of xe2x80x94NHxe2x80x94C(NR9R0)xe2x95x90NR11, xe2x80x94NHxe2x80x94C(R9)xe2x95x90NR, xe2x80x94C(NR9R10)xe2x95x90NR11 and piperidinyl; where R9, R10 and R11 are independently selected from the group consisting of H, C1-6alkyl, aryl-C1-3alkyl and aryl; or where two of the R9, R10 or R11 substituents form a cyclic ring containing (CH2)p, where p=2-5;
R1 is H;
R2 is selected from the group consisting of xe2x80x94SOm-aryl, xe2x80x94SOmxe2x80x94C1-10alkyl and xe2x80x94SOm-heteroaryl, where m=1-2;
R3 is selected from the group consisting of H, C1-8alkyl, aryl, C1-8alkylaryl and heteroaryl;
and all pharmaceutically-acceptable stereoisomers, salts, hydrates, solvates and prodrug derivatives thereof.
It is understood that the xe2x80x9cAxe2x80x9d substituents can be incorporated in the compounds of the invention in the order written above or in the reverse order. For example, a suitable xe2x80x9cAxe2x80x9d substituent is C0-8alkyl-Oxe2x80x94C0-8alkyl-COxe2x80x94NR5-C0-8alkyl. It is understood that the xe2x80x9cZxe2x80x9d substituent can be positioned to the right or to the left of this sequence.
Preferred xe2x80x9cYxe2x80x9d substituents are xe2x80x94COOH and xe2x80x94COOR4, more preferably xe2x80x94COOH. R4 is preferably C1-10alkyl.
Preferred xe2x80x9cAxe2x80x9d substituents are selected from the group consisting of C0-8alkyl-NR5xe2x80x94COxe2x80x94C0-8alkyl, C0-8alkyl-COxe2x80x94NR5xe2x80x94C0-8alkyl, C1-8alkyl-NR5xe2x80x94COxe2x80x94C1-8alkyl-NR5xe2x80x94COxe2x80x94C0-8alkyl, C0-8alkyl-NR5xe2x80x94COxe2x80x94C0-8alkyl-COxe2x80x94NR5-C0-8alkyl, C1-8alkyl-COxe2x80x94NR5-C1-8alkyl-NR5xe2x80x94COxe2x80x94C0-8alkyl, C0-8alkyl-COxe2x80x94NR5xe2x80x94C0-8alkyl-COxe2x80x94NR5xe2x80x94C0-8alkyl, C0-8alkyl-COxe2x80x94C1-8alkyl-COxe2x80x94NR5-C0-8alkyl, C0-8alkyl-Oxe2x80x94C2-8alkyl-NR5xe2x80x94COxe2x80x94C0-8alkyl, C0-8alkyl-Oxe2x80x94C0-8alkyl-COxe2x80x94NR5-C0-8alkyl, C0-8alkyl-Oxe2x80x94C2-8alkyl-Oxe2x80x94C0-8alkyl-COxe2x80x94NR5xe2x80x94C0-8alkyl, C0-8alkyl-Sxe2x80x94C2-8alkyl-NR5xe2x80x94COxe2x80x94C0-8alkyl, C0-8alkyl-S(On)-C2-8alkyl-NR5xe2x80x94COxe2x80x94C0-8alkyl, C0-8alkyl-Sxe2x80x94C0-8alkyl-COxe2x80x94NR5xe2x80x94C0-8alkyl, C0-8alkyl-S(On)-C1-8alkyl-COxe2x80x94NR5xe2x80x94C0-8alkyl, C0-8alkyl-NR5xe2x80x94COxe2x80x94C0-8alkyl-Sxe2x80x94C0-8alkyl, C0-8alkyl-NR5xe2x80x94COxe2x80x94C1-8alkyl-S(On)xe2x80x94C0-8alkyl, C0-8alkyl-COxe2x80x94NR5-C2-8alkyl-Sxe2x80x94C0-8alkyl, C0-8alkyl-CO-NR5-C2-8alkyl-S(On)-C0-8alkyl, C0-8alkyl-NR5-C0-8alkyl-CO2xe2x80x94C0-8alkyl, C0-8alkyl-NR5-C0-8alkyl-CSxe2x80x94Oxe2x80x94C0-8alkyl, C0-8alkyl-NR5-C0-8alkyl-COxe2x80x94NRxe2x80x94C0-8alkyl, C0-8alkyl-NR5xe2x80x94C0-8alkyl-CSxe2x80x94NR5-C0-8alkyl, C0-8alkyl-Oxe2x80x94C0-8alkyl-CO2xe2x80x94C0-8alkyl, C0-8alkyl-Oxe2x80x94C0-8alkyl-CSxe2x80x94C0-8alkyl; C0-8alkyl-SiR7R8-C0-8alkyl, C0-8alkyl-SiR7R8-C0-8alkyl-NR6xe2x80x94COxe2x80x94C0-8alkyl and C0-8alkyl-SiR7R8-C0-8alkyl-COxe2x80x94NR6xe2x80x94C0-8alkyl.
More preferred xe2x80x9cAxe2x80x9d substituents are selected from the group consisting of C0-8alkyl-COxe2x80x94NR5xe2x80x94C0-8alkyl, C0-8alkyl-Oxe2x80x94C0-8alkyl-COxe2x80x94NR5xe2x80x94C0-8alkyl, C0-8alkyl-Oxe2x80x94C2-8alkyl-Oxe2x80x94C0-8alkyl-COxe2x80x94NR5xe2x80x94C0-8alkyl, C0-8alkyl-Sxe2x80x94C0-8alkyl-COxe2x80x94NR5xe2x80x94C0-8alkyl, C0-8alkyl-S(On)-C1-8alkyl-COxe2x80x94NR5xe2x80x94C0-8alkyl, C0-8alkyl-NR5xe2x80x94COxe2x80x94C0-8alkyl-Sxe2x80x94C0-8alkyl, C0-8alkyl-NR5xe2x80x94COxe2x80x94C1-8alkyl-S(On)xe2x80x94C0-8alkyl, C0-8alkyl-NR5xe2x80x94C0-8alkyl-CO2xe2x80x94C0-8alkyl, C0-8alkyl-NR5xe2x80x94C0-8alkyl-COxe2x80x94NR5xe2x80x94C0-8alkyl, C0-8alkyl-Oxe2x80x94C0-8alkyl-CO2-C0-8alkyl.
Preferably, xe2x80x9cZxe2x80x9d is xe2x80x94NHxe2x80x94C(NR9R10)xe2x95x90NR11. Preferably R9, R10 and R11 are independently selected from the group consisting of H and C1-6alkyl. More preferably, R9, R10 and R11 are H.
Preferred R2 substituents are xe2x80x94SO2-aryl and xe2x80x94SO2xe2x80x94C1-10alkyl. More preferably, R2 is xe2x80x94SO2-aryl.
Preferred R3 substituents are H and C1-8alkyl. More preferably, R3 is H.
Preferred compounds and subgroups of compounds may be selected from any combination of the formulas presented in this specification with one or more of the preferred groupings of substituents at a particular location.
Other preferred compounds of the present invention are shown but not limited to the following list of compounds which have the structure: 
This invention also encompasses prodrug derivatives of the compounds contained herein. The term xe2x80x9cprodrugxe2x80x9d refers to a pharmacologically-inactive derivative of a parent drug molecule that requires biotransformation, either spontaneous or enzymatic, within the organism to release the active drug. Prodrugs are variations or derivatives of the compounds of this invention which have metabolically cleavable groups and become, by solvolysis under physiological conditions, or by enzymatic degradation, compounds which are pharmaceutically active in vivo. Prodrug compounds of this invention may be called single, double, triple etc., depending on the number of biotransformation steps required to release the active drug within the organism, and indicating the number of functionalities present in a precursor-type form. Prodrug forms often offer advantages of solubility, tissue compatibility, or delayed release in the mammalian organism (see, Bundgard, H., Design of Prodruqs, pp. 7-9, 21-24, Elsevier, Amsterdam 1985 and Silverman, R. B., The Organic Chemistry of Drug Design and Drug Action, pp. 352-401, Academic Press, San Diego, Calif., 1992). Prodrugs commonly known in the art include acid derivatives well known to practitioners of the art, such as, for example, esters prepared by reaction of the parent acids with a suitable alcohol, or amides prepared by reaction of the parent acid compound with an amine, or basic groups reacted to form an acylated base derivative. Moreover, the prodrug derivatives of this invention may be combined with other features herein taught to enhance bioavailability.
Preparation of Compounds
The compounds of the present invention may be synthesized by either solid or liquid phase methods described and referenced in standard textbooks, or by a combination of both methods. These methods are well known in the art. See, Bodanszky, M., in xe2x80x9cThe Principles of Peptide Synthesisxe2x80x9d, Hafner, K., Rees, C. W., Trost, B. M., Lehn, J.-M., Schleyer, P. v-R., Zahradnik, R., Eds., Springer-Verlag, Berlin, 1984. Starting materials are commercially available reagents and reactions are carried out in standard laboratory glassware and reaction vessels under reaction conditions of standard temperature and pressure, except where otherwise indicated.
The starting materials used in any of these methods are commercially available from chemical vendors such as Aldrich, Sigma, Nova Biochemicals, Bachem Biosciences, and the like, or may be readily synthesized by known procedures.
During the synthesis of these compounds, the functional groups of the amino acid derivatives used in these methods are protected by blocking groups to prevent cross reaction during the coupling procedure. Examples of suitable blocking groups and their use are described in xe2x80x9cThe Peptides: Analysis, Synthesis, Biologyxe2x80x9d, Academic Press, Vol. 3 (Gross, E. and Meienhofer, J., Eds., 1981) and Vol. 9 (, S. and., Eds., 1987), the disclosures of which are incorporated herein by reference.
Nine exemplary synthesis schemes are outlined below, and the specific syntheses are described in the Examples. The reaction products are isolated and purified by conventional methods, typically by solvent extraction into a compatible solvent. The products may be further purified by column chromatography or other appropriate methods. 
The preparation of the carbamate containing compound 107 typifies the construction of compounds containing this linkage (Scheme 3-A). In the first step, a suitable protected amino alcohol (illustrated with analog 100) is allowed to react with excess phosgene to provide the intermediate chloroformate (101). This material is condensed with the differentially protected diamino propionate derivative 103, thus producing the carbamate linked intermediate (104). The CBz group on the terminal amine is then removed with palladium and hydrogen (105) and the formed amine is reacted with N,Nxe2x80x2-bis(tert-butoxycarbonyl)-S-methoxy-isothiourea producing the protected guanidine derivative (106). The compound is completely deprotected with neat TFA at room temperature and then subjected to a salt exchange with HCI providing the desired amino acids (107). Compounds containing a urea linkage (113) are prepared in an analogous manner starting from differentially protected diamines (Scheme 3-B).
Compounds that contain an amine linkage (118) can be prepared in a similar fashion (Scheme 3-A). Protected bromo-amine 114 (prepared from alcohol 113 by treatment with CBr4 and Ph3P) is allowed to react with amine 103 in the presence of K2CO3 which forms adduct 115. Subjection of protected derivative 115 to the same sequence of reactions outlined for the transformation of 104 into 107 affords the desired amine containing compound 118.
Compositions and Formulations
The compounds of this invention may be isolated as the free acid or base or converted to salts of various inorganic and organic acids and bases. Such salts are within the scope of this invention. Non-toxic and physiologically-compatible salts are particularly useful although other less desirable salts may have use in the processes of isolation and purification.
A number of methods are useful for the preparation of the salts described above and are known to those skilled in the art. For example, reaction of the free acid or free base form of a compound of the structures recited above with one or more molar equivalents of the desired acid or base in a solvent or solvent mixture in which the salt is insoluble, or in a solvent like water after which the solvent is removed by evaporation, distillation or freeze drying. Alternatively, the free acid or base form of the product may be passed over an ion exchange resin to form the desired salt or one salt form of the product may be converted to another using the same general process.
Diagnostic applications of the compounds of this invention will typically utilize formulations such as solution or suspension. In the management of thrombotic disorders the compounds of this invention may be utilized in compositions such as tablets, capsules or elixirs for oral administration, suppositories, sterile solutions or suspensions or injectable administration, and the like, or incorporated into shaped articles. Subjects in need of treatment (typically mammalian) using the compounds of this invention can be administered dosages that will provide optimal efficacy. The dose and method of administration will vary from subject to subject and be dependent upon such factors as the type of mammal being treated, its sex, weight, diet, concurrent medication, overall clinical condition, the particular compounds employed, the specific use for which these compounds are employed, and other factors which those skilled in the medical arts will recognize.
Formulations of the compounds of this invention are prepared for storage or administration by mixing the compound having a desired degree of purity with physiologically acceptable carriers, excipients, stabilizers etc., and may be provided in sustained release or timed release formulations. Acceptable carriers or diluents for therapeutic use are well known in the pharmaceutical field, and are described, for example, in Remington""s Pharmaceutical Sciences, Mack Publishing Co., (A. R. Gennaro edit. 1985). Such materials are nontoxic to the recipients at the dosages and concentrations employed, and include buffers such as phosphate, citrate, acetate and other organic acid salts, antioxidants such as ascorbic acid, low molecular weight (less than about ten residues) peptides such as polyarginine, proteins, such as serum albumin, gelatin, or immunoglobulins, hydrophilic polymers such as polyvinylpyrrolidinone, amino acids such as glycine, glutamic acid, aspartic acid, or arginine, monosaccharides, disaccharides, and other carbohydrates including cellulose or its derivatives, glucose, mannose or dextrins, chelating agents such as EDTA, sugar alcohols such as mannitol or sorbitol, counterions such as sodium and/or nonionic surfactants such as Tween, Pluronics or polyethyleneglycol.
Dosage formulations of the compounds of this invention to be used for therapeutic administration must be sterile. Sterility is readily accomplished by filtration through sterile membranes such as 0.2 micron membranes, or by other conventional methods. Formulations typically will be stored in lyophilized form or as an aqueous solution. The pH of the preparations of this invention typically will be between 3 and 11, more preferably from 5 to 9 and most preferably from 7 to 8. It will be understood that use of certain of the foregoing excipients, carriers, or stabilizers will result in the formation of cyclic polypeptide salts. While the preferred route of administration is by injection, other methods of administration are also anticipated such as intravenously (bolus and/or infusion), subcutaneously, intramuscularly, colonically, rectally, nasally or intraperitoneally, employing a variety of dosage forms such as suppositories, implanted pellets or small cylinders, aerosols, oral dosage formulations and topical formulations such as ointments, drops and dermal patches. The compounds of this invention are desirably incorporated into shaped articles such as implants which may employ inert materials such as biodegradable polymers or synthetic silicones, for example, Silastic, silicone rubber or other polymers commercially available.
The compounds of this invention may also be administered in the form of liposome delivery systems, such as small unilamellar vesicles, large unilamellar vesicles and multilamellar vesicles. Liposomes can be formed from a variety of lipids, such as cholesterol, stearylamine or phosphatidylcholines.
The compounds of this invention may also be delivered by the use of antibodies, antibody fragments, growth factors, hormones, or other targeting moieties, to which the compound molecules are coupled. The compounds of this invention may also be coupled with suitable polymers as targetable drug carriers. Such polymers can include polyvinylpyrrolidinone, pyran copolymer, polyhydroxy-propyl-methacrylamide-phenol, polyhydroxyethyl-aspartamide-phenol, or polyethyleneoxide-polylysine substituted with palmitoyl residues. Furthermore, inhibitors of this invention may be coupled to a class of biodegradable polymers useful in achieving controlled release of a drug, for example polylactic acid, polyglycolic acid, copolymers of polylactic and polyglycolic acid, polyepsilon caprolactone, polyhydroxy butyric acid, polyorthoesters, polyacetals, polydihydropyrans, polycyanoacrylates and cross linked or amphipathic block copolymers of hydrogels. Polymers and semipermeable polymer matrices may be formed into shaped articles, such as valves, stents, tubing, prostheses and the like.
Therapeutic compound liquid formulations generally are placed into a container having a sterile access port, for example, an intravenous solution bag or vial having a stopper pierceable by hypodermic injection needle.
Therapeutically-effective dosages may be determined by either in vitro or in vivo methods. For each particular compound of the present invention, individual determinations may be made to determine the optimal dosage required. The range of therapeutically-effective dosages will naturally be influenced by the route of administration, the therapeutic objectives, and the condition of the patient. For injection by hypodermic needle, it may be assumed the dosage is delivered into the bodys fluids. For other routes of administration, the absorption efficiency must be individually determined for each inhibitor by methods well known in pharmacology. Accordingly, it may be necessary for the therapist to titer the dosage and modify the route of administration as required to obtain the optimal therapeutic effect. The determination of effective dosage levels, that is, the dosage levels necessary to achieve the desired result, will be within the ambit of one skilled in the art. Typically, applications of compound are commenced at lower dosage levels, with dosage levels being increased until the desired effect is achieved.
A typical dosage might range from about 0.001 mg/kg to about 1000 mg/kg, preferably from about 0.01 mg/kg to about 100 mg/kg, and more preferably from about 0.10 mg/kg to about 20 mg/kg. Advantageously, the compounds of this invention may be administered several times daily, and other dosage regimens may also be useful.
Typically, about 0.5 to 500 mg of a compound or mixture of compounds of this invention, as the free acid or base form or as a pharmaceutically-acceptable salt, is compounded with a physiologically acceptable vehicle, carrier, excipient, binder, preservative, stabilizer, dye, flavor etc., as called for by accepted pharmaceutical practice. The amount of active ingredient in these compositions is such that a suitable dosage in the range indicated is obtained.
Typical adjuvants which may be incorporated into tablets, capsules and the like are a binder such as acacia, corn starch or gelatin, and excipient such as microcrystalline cellulose, a disintegrating agent like corn starch or alginic acid, a lubricant such as magnesium stearate, a sweetening agent such as sucrose or lactose, or a flavoring agent. When a dosage form is a capsule, in addition to the above materials it may also contain a liquid carrier such as water, saline, or a fatty oil. Other materials of various types may be used as coatings or as modifiers of the physical form of the dosage unit. Sterile compositions for injection can be formulated according to conventional pharmaceutical practice. For example, dissolution or suspension of the active compound in a vehicle such as an oil or a synthetic fatty vehicle like ethyl oleate, or into a liposome may be desired. Buffers, preservatives, antioxidants and the like can be incorporated according to accepted pharmaceutical practice.
In practicing the methods of this invention, the compounds of this invention may be used alone, or in combination with other therapeutic or diagnostic agents. In certain preferred embodiments, the compounds of this invention may be coadministered along with other compounds typically prescribed for these conditions according to generally accepted medical practice, such as anticoagulant agents, thrombolytic agents, or other antithrombotics, including platelet aggregation inhibitors, tissue plasminogen activators, urokinase, prourokinase, streptokinase, heparin, aspirin, or warfarin. The compounds of this invention can be utilized in vivo, ordinarily in mammals such as primates, (e.g. humans), sheep, horses, cattle, pigs, dogs, cats, rats and mice, or in vitro.
With respect to the coronary arterial vasculature, abnormal thrombus formation characterizes the rupture of an established atherosclerotic plaque which is the major cause of acute myocardial infarction and unstable angina, as well as also characterizing the occlusive coronary thrombus formation resulting from either thrombolytic therapy or percutaneous transluminal coronary angioplasty (PTCA).
The compounds of the present invention preferably have an IC50 of less than about 2.0 xcexcM, more preferably less than about 1.0 xcexcM, and most preferably less than about 200 nM, as measured by one or more of the assays described herein. These compounds, selected and used as disclosed herein, are believed to be useful for preventing or treating a condition characterized by undesired thrombosis, such as, by way of illustration and not limitation, (a) the treatment or prevention of any thrombotically mediated acute coronary syndrome including myocardial infarction, unstable angina, refractory angina, occlusive coronary thrombus occurring post-thrombolytic therapy or post-coronary angioplasty, (b) the treatment or prevention of any thrombotically mediated cerebrovascular syndrome including embolic stroke, thrombotic stroke or transient ischemic attacks, (c) the treatment or prevention of any thrombotic syndrome occurring in the venous system including deep venous thrombosis or pulmonary embolus occurring either spontaneously or in the setting of malignancy, surgery or trauma, thrombotic thrombocytopenic purpura, thromboangiitis obliterans, or thrombotic disease associated with heparin induced thrombocytopenia, (e) the treatment or prevention of thrombotic complications associated with extracorporeal circulation (e.g. renal dialysis, cardiopulmonary bypass or other oxygenation procedure, plasmapheresis), (f) coagulopathy and disseminated intravascular coagulation (g) the treatment or prevention of thrombotic complications associated with instrumentation (e.g. cardiac or other intravascular catheterization, intra-aortic balloon pump, coronary stent or cardiac valve), (h) those involved with the fitting of prosthetic devices, (i) vascularization of solid tumors and (j) retinopathy.
Without further description, it is believed that one of ordinary skill in the art can, using the preceding description and the following illustrative examples, make and utilize the compounds of the present invention and practice the claimed methods. The following working examples therefore, specifically point out preferred embodiments of the present invention, and are not to be construed as limiting in any way the remainder of the disclosure.